兰州市娃娃菜褐腐病菌生物学特性及融合群鉴定

doi:10.11924/j.issn.1000-6850.casb18040138

中国农学通报 ›› 2019, Vol. 35 ›› Issue (14): 129-137.doi: 10.11924/j.issn.1000-6850.casb18040138

所属专题：园艺

兰州市娃娃菜褐腐病菌生物学特性及融合群鉴定

何苏琴¹, 文朝慧², 白滨³, 荆卓琼⁴

1.甘肃省农业科学院植物保护研究所;2.甘肃出入境检验检疫局;3.甘肃省农业科学院农业质量标准与检测技术研究所;4.甘肃省农业科学院植物保护研究所

收稿日期:2018-04-28 修回日期:2019-04-18 接受日期:2018-07-03 出版日期:2019-05-15 发布日期:2019-05-15
通讯作者: 何苏琴
基金资助:
甘肃省农业科技创新专项“兰州百合贮藏期病害防控技术研究”(GNCX-2014-34)；甘肃出入境检验检疫局科技计划项目“防控农业有害生物随外繁制种入侵我省的技术体系构建与应用”(2015GK001)；甘肃省农业科学院农业科技创新专项“生物防治技术研究与应用”(2017GAAS23)。

Biological Characteristics and Anastomosis Groups of Rhizoctonia solani Associated with Chinese Cabbage Brown Rot in Lanzhou

Received:2018-04-28 Revised:2019-04-18 Accepted:2018-07-03 Online:2019-05-15 Published:2019-05-15

摘要/Abstract

摘要： 为给病害科学防治提供基础信息，对分离自兰州市红古区和永登县武胜驿镇罹病娃娃菜病株上的8 株立枯丝核菌菌株的形态、生长温度、致病性、对杀菌剂的敏感性等生物学特性进行研究，对其菌丝融合群归属进行分子生物学鉴定。采用温度梯度法测定病菌适宜生长温度，同时观察试验菌株微菌核产生情况；采用番红O和KOH染色法观测试验菌株细胞核数目；采用离体叶片接种法测定试验菌株的致病力；采用含药平板法测定试验菌株对5 种杀菌剂的敏感性；以试验菌株的5.8S rDNA-ITS 区序列构建系统发育树，进行融合群的分子生物学鉴定。结果表明，除菌株Rh-5 适宜生长温度为15~20℃外，其余试验菌株适宜生长温度均为25℃。试验菌株在5~30℃培养均可形成微菌核。菌株Rh-1~Rh-8 的细胞核数目分别为3~6、6~14、3~18、4~15、3~10、4~10、3~10、4~8 个。接种Rh-2~Rh-6 的娃娃菜离体叶片在20℃保湿培养2 天内发病显症，7 天发病率达75%~100%；接种Rh-1~Rh-8 的娃娃菜离体叶片在25℃保湿培养1~3 天内发病显症，7 天发病率达25%~100%；接种Rh-1、Rh-7 和Rh-8 的娃娃菜离体叶片在28℃保湿培养2~3 天发病显症，7 天发病率达100%。在试验浓度下，8 个试验菌株对5 种杀菌剂的敏感性由大到小依次为多菌灵、代森锰锌、速克灵、三唑酮、苯醚甲环唑。除Rh-1 与Rhizoctonia solani AG 4 HG-II 聚为一枝外，其余试验菌株（Rh-6 除外，因Rh-6 测序未果）均与R. solani AG 2-1 聚为一枝。依据试验菌株的5.8S rDNA-ITS 区序列分析所得结果，引起兰州市娃娃菜褐腐病的立枯丝核菌包括AG 2-1 和AG 4HG-II 融合群。试验菌株在菌落形态、细胞核数目、适宜生长温度、生长速度、侵染温度、致病性及对杀菌剂的敏感性等方面呈现出丰富的多样性特征。在试验浓度下，多菌灵和代森锰锌对试验菌株的抑制率≥80%，可作为田间病害防治试验的推荐药剂。

关键词: 海滨雀稗, 海滨雀稗, 分子标记, EST-SSR, 遗传多样性

Abstract: To provide information for the disease control, 8 strains of Rhizoctonia solani isolated from diseased mini Chinese cabbage in Honggu and Yongdeng of Lanzhou were used for testing morphology, cardinal temperature, pathogenicity, fungicide sensibility, and anastomosis group identification. The optimal growth temperatures were evaluated by temperature gradient method, and microsclerotia forming status was observed in the meantime. Numbers of nuclei were observed by staining with safranin O and KOH. The pathogenicity was determined by inoculation to detached leaves. Fungicide sensitivity of R. solani strains was assessed by amendment plate method, autoclaved PDA media were amended prior to pouring with 5 different fungicides. Phylogenetic tree was established based on rDNA- ITS (ITS1- 5.8S- ITS2) sequences of test strains. The results showed that: the optimal growth temperature of all other strains was 25℃, except Rh-5, whose was 15-20℃. All tested strains could produce microsclerotia cultured on PDA plate at 5-30℃. Numbers of nuclei of strain Rh-1- Rh-8 was 3-6, 6-14, 3-18, 4-15, 3-10, 4-10, 3-10 and 4-8, respectively. Detached leaves of mini Chinese cabbage inoculated by strain Rh-2-Rh-6, infected in 2 d at 20℃, had the incidence of 75%-100% in 7 d after inoculation; the leaves inoculated by strain Rh-1-Rh-8, infected in 1-3 d at 25℃, had the incidence of 25%- 100% in 7 d after inoculation; the leaves inoculated by strain Rh-1, Rh-7 and Rh-8, infected in 2-3 d at 28℃, had the incidence of 100% in 7 d after inoculation. Under test concentrations, the sensitivity of the eight tested strains of R. solani to five fungicides was successively: carbendazim> mancozeb> procymidone> triadimefon> difenoconazole. Rh- 1 aligned with R. solani AG 4 HG- II, other test strains (except Rh- 6, since sequencing failure) aligned with R. solani AG 2-1. According to rDNA-ITS (ITS1-5.8S-ITS2) sequences analysis of the test strains, the pathogens causing mini Chinese cabbage brown rot in Lanzhou include R. solani AG 2-1 and AG 4 HG-II. Tested strains of R. solani show the diversified characteristics in colony morphology, numbers of nuclei, optimal growth temperatures, growth speeds, infection temperatures, pathogenicity and fungicide sensitivity. At the tested concentrations, all strains show higher sensitivity to carbendazim and mancozeb (inhibition rate≥ 80%), suggesting that these two fungicides could be used in field trials on control of mini Chinese cabbage brown rot disease.

何苏琴,文朝慧,白滨,荆卓琼. 兰州市娃娃菜褐腐病菌生物学特性及融合群鉴定[J]. 中国农学通报, 2019, 35(14): 129-137.

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兰州市娃娃菜褐腐病菌生物学特性及融合群鉴定

Biological Characteristics and Anastomosis Groups of Rhizoctonia solani Associated with Chinese Cabbage Brown Rot in Lanzhou

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